Matsuzaki H, Araki H, Oshima Y
Department of Fermentation Technology, Faculty of Engineering, Osaka University, Japan.
Mol Cell Biol. 1988 Feb;8(2):955-62. doi: 10.1128/mcb.8.2.955-962.1988.
A circular DNA plasmid, pSR1, isolated from Zygosaccharomyces rouxii has a pair of inverted repeats consisting of completely homologous 959-base pair (bp) sequences. Intramolecular recombination occurs frequently at the inverted repeats in cells of Saccharomyces cerevisiae, as well as in Z. rouxii, and is catalyzed by a protein encoded by the R gene of its own genome. The recombination is, however, independent of the RAD52 gene of the host genome. A site for initiation of the intramolecular recombination in the S. cerevisiae host was delimited into, at most, a 58-bp region in the inverted repeats by using mutant plasmids created by linker insertion. The 58-bp region contains a pair with 14-bp dyad symmetry separated by a 3-bp spacer sequence. The recombination initiated at this site was accompanied by a high frequency of gene conversion (3 to 50% of the plasmid clones examined). Heterogeneity created by the linker insertion or by a deletion (at most 153 bp so far tested) at any place on the inverted repeats was converted to a homologous combination by the gene conversion, even in the rad52-1 mutant host. A mechanism implying branch migration coupled with DNA replication is discussed.
从鲁氏接合酵母中分离出的环状DNA质粒pSR1有一对由完全同源的959碱基对(bp)序列组成的反向重复序列。分子内重组在酿酒酵母细胞以及鲁氏接合酵母细胞的反向重复序列处频繁发生,并由其自身基因组的R基因编码的一种蛋白质催化。然而,这种重组不依赖于宿主基因组的RAD52基因。通过使用由接头插入产生的突变质粒,在酿酒酵母宿主中分子内重组的起始位点被限定在反向重复序列中最多58 bp的区域内。该58 bp区域包含一对由3 bp间隔序列隔开的具有14 bp二元对称的序列。在此位点起始的重组伴随着高频的基因转换(在所检测的质粒克隆中有3%至50%)。即使在rad52-1突变宿主中,由接头插入或反向重复序列上任何位置的缺失(迄今为止测试的最大缺失为153 bp)所产生的异质性也通过基因转换转变为同源组合。讨论了一种暗示分支迁移与DNA复制耦合的机制。